All AFQ-056 embryos were randomly allocated to a treatment method 
group. Data had been analyzed with a 1-way examination of variance and Tukey’s minimum important check employing GraphPad Prism six computer software. p<0.05 was considered significant.After activation, SCNT embryos were treated with various concentrations of Scriptaid (0, 100, 300, and 500 nM) for 20 h and their in vitro development was examined. The percentage of Fig 1. Effect of treatment with Scriptaid at various concentrations (a and b) and for various amounts of time (c, d, and e) on the development of porcine somatic cell nuclear transfer embryos to the blastocyst stage. The numbers of embryos examined in each experimental group are shown in the bars. indicates P<0.05 compared with the control group. Values are the mean (standard deviation of the mean) of four independent experiments. In E, embryos were not treated or were treated with 300 nM Scriptaid for 20 h. Ctrl: no treatment SCR: Scriptaid treatment. Magnification, 100. embryos that developed to the blastocyst stage was significantly higher among embryos treated with 300 or 500 nM Scriptaid than among those treated with 100 nM Scriptaid and non-treated embryos (300 nM, 23.50% 500 nM, 25.78% 100 nM, 16.48% non-treated, 15.26% Fig 1A). However, the percentage of embryos that developed to the blastocyst stage did not significantly differ between those treated with 300 nM Scriptaid and those treated with 500 nM Scriptaid. Similar results were observed for the total number of cells per blastocyst (Fig 1B), which was significantly higher among embryos treated with 300 nM or 500 nM Scriptaid than among those treated with 100 nM Scriptaid and non-treated embryos. Based on these results, 300 nM was chosen as the optimal concentration of Scriptaid. Next, the effects of treatment with 300 nM Scriptaid for various amounts of time (0, 10, and 20 h) on the in vitro development of SCNT embryos were analyzed. The percentage of SCNT embryos that developed to the blastocyst stage was significantly higher among those treated with Fig 2. Laser scanning confocal microscopy images of nuclei and fragmented DNA (00, a and b) and relative mRNA expression levels of Bcl-xL, Bax, and Caspase3 (Cas3) (c) in porcine blastocysts after 7 days of in vitro culture, with or without Scriptaid treatment. The numbers of embryos examined in each experimental group are shown in the bars. indicates P<0.05 compared with the control group. Values are the mean (standard deviation of the mean) of four independent experiments. Ctrl: no treatment SCR: Scriptaid treatment.Scriptaid for 20 h (24.90%) than among those treated with Scriptaid for 0 h (15.19%) or 10 h (17.02%) (Fig 1C and 1E). The total 1674587number of cells per blastocyst was significantly higher among Scriptaid-treated embryos (10 or 20 h) than among non-treated embryos (Fig 1D).DNA fragments generated by apoptotic nicking of genomic DNA were measured in individual embryos using the TUNEL assay.